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1. WO2020113274 - RANK ANTAGONISTS AND USES THEREFOR

Note: Text based on automatic Optical Character Recognition processes. Please use the PDF version for legal matters

[ EN ]

WHAT IS CLAIMED IS:

1. An RANK antagonist antigen-binding molecule comprising :

(1) a heavy chain variable region (VH) comprising a VHCDR1 amino acid sequence set forth in SEQ ID NO: 3, a VHCDR2 amino acid sequence set forth in SEQ ID NO:4, and a VHCDR3 amino acid sequence set forth in SEQ ID NO: 5, and a light chain variable region (\ ) comprising a VLCDR1 amino acid sequence set forth in SEQ ID NO: 6, a VLCDR2 amino acid sequence set forth in SEQ ID NO: 7, and a VLCDR3 amino acid sequence set forth in SEQ ID NO: 8;

(2) a V that comprises the amino acid sequence set forth in SEQ ID NO: l, and a VL that comprises the amino acid sequence set forth in SEQ ID NO: 2;

(3) a V with at least 90% (including at least 91% to 99% and all integer percentages therebetween) sequence identity to the amino acid sequence of SEQ ID NO: 1, and a VL with at least 90% (including at least 91% to 99% and all integer percentages therebetween) sequence identity to the amino acid sequence of SEQ ID NO: 2;

(4) a V with at least 90% (including at least 91% to 99% and all integer percentages therebetween) sequence identity to the amino acid sequence of a framework region other than each CDR in the amino acid sequence of SEQ ID NO: l, and a VL with at least 90% (including at least 91% to 99% and all integer percentages therebetween) sequence identity to the amino acid sequence of a framework region other than each CDR in the amino acid sequence of SEQ ID NO: 2; or

(5) a V that comprises an amino acid sequence comprising a deletion, substitution or addition of one or more {e.g. , 1, 2, 3, 4 or 5) amino acids in the sequence of a framework region other than at each CDR in the amino acid sequence of SEQ ID NO: 1, and a VL that comprises an amino acid sequence comprising a deletion, substitution or addition of one or more {e.g. , 1, 2, 3, 4 or 5) amino acids in the sequence of a framework region other than at each CDR in the amino acid sequence of SEQ ID NO: 2.

2. The antigen-binding molecule of claim 1, wherein the antigen-binding molecule is in isolated, purified, synthetic or recombinant form.

3. The antigen-binding molecule of claim 1 or claim 2, wherein the antigen binding molecule is monovalent {e.g. , Fab, scFab, Fab', scFv, one-armed antibodies, etc.).

4. The antigen-binding molecules of any one or more of claims 1 to 3, having any one or more of the following activities: (a) inhibits binding of RANKL to

RANK; (b) inhibits RANK activation; (c) inhibits downstream RANK-mediated molecular signaling {e.g., RANK recruitment of TRAF proteins) ; (d) inhibits RANK multimerization; (e) reduces osteoclast differentiation; (f) decreases osteoclast activation; (g) reduces osteoclast survival ; (h) inhibits bone loss and increase bone density; (i) inhibits immunosuppressive activity of myeloid cells or other immune cells in a tumor microenvironment (TME) ; and (j) inhibits proliferation, migration, survival and/or morphogenesis of tumor cells {e.g. , breast cancer cells including triple negative breast cancer (TN BC) cells and BRCA-1 mutation positive breast cancer cells, non-small cell lung cancer (NSCLC) cells and renal cell carcinoma (RCC) cells).

5. The antigen-binding molecules of any one or more of claims 1 to 3, wherein the antigen-binding molecule is contained in a delivery vehicle (e.g., a liposome, a nanoparticle, a microparticle, a dendrimer or a cyclodextrin).

6. An isolated polynucleotide comprising a nucleic acid sequence encoding the RANK antagonist antigen-binding molecule of any one of claims 1 to 4.

7. A construct comprising a nucleic acid sequence encoding the RANK antagonist antigen-binding molecule of any one of claims 1 to 4 in operable connection with one or more control sequences.

8. The construct of claim 7, which is in the form of an expression construct (e.g. , a plasmid, cosmid, phage, or virus).

9. A host cell that contain the construct of claim 8.

10. A pharmaceutical composition comprising the RANK antagonist antigen-binding molecule of any one of claims 1 to 4 and a pharmaceutically acceptable carrier.

11. The composition of claim 10, further comprising at least one ancillary agent selected from a bone anti-resorptive agent (e.g. , anabolism enhancers, in particular selected from the group consisting of parathyroid hormone, BMP2, vitamin D, anti-inflammatory agents; and catabolism inhibitors, in particular selected from the group consisting of bisphosphonates, cathepsin K inhibitors, p38 inhibitors, JNK inhibitors, IKK inhibitors, NF— KB inhibitors, calcineurin inhibitors, NFAT inhibitors, PI3K inhibitors) and a chemotherapeutic agent (e.g., antiproliferative/antineoplastic drugs, cytostatic agents, agents that inhibit cancer cell invasion, inhibitors of growth factor function, anti-angiogenic agents, vascular damaging agents, etc.) or an

immunotherapeutic agent (e.g. , cytokines, cytokine-expressing cells, antibodies, etc.).

12. A method for inhibiting binding of RANKL to a RANK-expressing cell, the method comprising contacting the RANK-expressing cell with the RANK antagonist antigen-binding molecule of any one of claims 1 to 4, to thereby inhibit binding of RANK to the RANK expressing cell.

13. A method for inhibiting activation of RANK on a RANK-expressing cell, the method comprising contacting the RANK-expressing cell with a RANK antagonist antigen-binding molecule of any one of claims 1 to 4, to thereby inhibit activation of RANK on the RANK expressing cell.

14. A method for inhibiting RANK-mediated molecular signaling (e.g., RANK recruitment of TRAF proteins) in a RANK-expressing cell, the method comprising contacting the RANK-expressing cell with a RANK antagonist antigen-binding molecule of any one of claims 1 to 4, to thereby inhibit RANK-mediated molecular signaling in the RANK expressing cell.

15. A method for inhibiting RANK multimerization in a RANK-expressing cell, the method comprising contacting the RANK-expressing cell with a RANK antagonist antigen-binding molecule described herein, to thereby inhibit RANK multimerization in the RANK expressing cell.

16. The method of any one of claim 12 to 15, wherein the RANK-expressing cell is selected from osteoclasts, immune cells such as antigen-presenting cells {e.g., monocytes and dendritic cells) and effector immune cells {e.g., T cells), hematopoietic precursors, and tumor cells such as breast cancer cells, prostate cancer cells, NSCLC cells and RCC cells.

17. A method for inhibiting differentiation, activation and/or survival of an osteoclast, the method comprising contacting the osteoclast with a RANK antagonist antigen-binding molecule of any one of claims 1 to 4, to thereby inhibit differentiation, activation and/or survival of the osteoclast.

18. A method for inhibiting immunosuppressive activity of an immune cell

(e.g., a myeloid cell or Treg), the method comprising contacting the immune cell with a RANK antagonist antigen-binding molecule of any one of claims 1 to 4, to thereby inhibit the immunosuppressive activity of the immune cell.

19. A method for inhibiting proliferation, survival or migration of a tumor cell, the method comprising contacting the tumor cell with a RANK antagonist antigen binding molecule of any one of claims 1 to 4, to thereby inhibit proliferation, survival or migration the tumor cell.

20. A method for treating or inhibiting the development of a condition associated with activation of the RANKL/RANK signaling pathway in a subject, the method comprising administering to the subject an effective amount of a RANK antagonist antigen-binding molecule of any one of claims 1 to 4, thereby treating or inhibiting the development of the condition.

21. The method of claim 20, wherein the condition associated with

RANKL/RANK signaling pathway activation is selected from an osteopenic disorder, a myopathy and a cancer.

22. A therapeutic combination comprising, consisting, or consisting essentially of the RANK antagonist antigen-binding molecule of any one of claim 1 to 4, and at least one anti-ICM antigen-binding molecule.

23. The therapeutic combination of claim 22, wherein the RANK antagonist antigen-binding molecule and the at least one anti-ICM antigen-binding molecule are in the form of a single composition (e.g., a mixture).

24. The therapeutic combination of claim 22, wherein the RANK antagonist antigen-binding molecule and the at least one anti-ICM antigen-binding molecule are in the form of discrete components in separate compositions.

25. The therapeutic combination of any one of claim 22 to 24, wherein the at least one anti-ICM antigen-binding molecule antagonizes an ICM selected from the group consisting of: programmed death 1 receptor (PD-1), programmed death ligand 1 (PD-L1), programmed death ligand 2 (PD-L2), cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4), A2A adenosine receptor (A2AR), A2B adenosine receptor (A2BR), B7-H3 (CD276), V-set domain-containing T-cell activation inhibitor 1 (VTCN1), B- and T-lymphocyte attenuator (BTLA), indoleamine 2,3-dioxygenase (IDO), killer-cell immunoglobulin-like receptor (KIR), lymphocyte activation gene-3 (LAG3), T cell immunoglobulin domain and mucin domain 3 (TIM-3), V-domain Ig suppressor of T cell

activation (VISTA), 5'-nucleotidase (CD73), tactile (CD96), poliovirus receptor (CD155), DNAX Accessory Molecule-1 (DNAM-1), poliovirus receptor-related 2 (CD112), cytotoxic and regulatory T-cell molecule (CRTAM), tumor necrosis factor receptor superfamily member 4 (TNFRS4; 0X40; CD134), tumor necrosis factor (ligand) superfamily, member 4 (TNFSF4; 0X40 ligand (OX40L), natural killer cell receptor 2B4 (CD244),

CD160, glucocorticoid-induced TNFR-related protein (GITR), glucocorticoid-induced TNFR-related protein ligand (GITRL), inducible co-stimulator (ICOS), galectin 9 (GAL-9), 4-1BB ligand (4-1BBL; CD137L), 4-1BB (4-1BB; CD137), CD70 (CD27 ligand (CD27L)), CD28, B7-1 (CD80), B7-2 (CD86), signal-regulatory protein (SIRP-1), integrin associated protein (IAP; CD47); B-lymphocyte activation marker (BLAST-1; CD48), natural killer cell receptor 2B4 (CD244); CD40, CD40 ligand (CD40L), herpesvirus entry mediator (HVEM), transmembrane and immunoglobulin domain containing 2 (TMIGD2), HERV-H LTR-associating 2 (HHLA2), vascular endothelial growth inhibitor (VEGI), tumor necrosis factor receptor superfamily member 25 (TNFRS25), inducible T-cell co-stimulator ligand (ICOLG; B7RP1) and T cell immunoreceptor with Ig and ITIM

(immunoreceptor tyrosine-based inhibition motif ) domains (TIGIT).

26. The therapeutic combination of claim 25, wherein the at least one anti-ICM antigen-binding molecule is selected from a PD-1 antagonist antigen-binding molecule, a PD-L1 antagonist antigen-binding molecule and a CTLA4 antagonist antigen-binding molecule.

27. The therapeutic combination of claim 25, wherein the at least one anti-ICM antigen-binding molecule comprises a PD-1 antagonist antigen-binding molecule.

28. The therapeutic combination of claim 25, wherein the at least one anti-ICM antigen-binding molecule comprises a PD-L1 antagonist antigen-binding molecule.

29. The therapeutic combination of claim 25, wherein the at least one anti- ICM antigen-binding molecule comprises a CTLA4 antagonist antigen-binding molecule.

30. The therapeutic combination of claim 25, wherein the at least one anti-ICM antigen-binding molecule comprises a PD-1 antagonist antigen-binding molecule and a PD-L1 antagonist antigen-binding molecule.

31. The therapeutic combination of claim 25, wherein the at least one anti- ICM antigen-binding molecule comprises a PD-1 antagonist antigen-binding molecule and a CTLA4 antagonist antigen-binding molecule.

32. The therapeutic combination of claim 25, wherein the at least one anti-ICM antigen-binding molecule comprises a PD-L1 antagonist antigen-binding molecule and a CTLA4 antagonist.

33. The therapeutic combination of claim 25, wherein the anti-ICM antigen binding molecule antagonizes an ICM that a Treg cell lacks expression of or expresses at a low level.

34. The therapeutic combination of claim 25, wherein the anti-ICM antigen-binding molecule antagonizes an ICM (e.g., PD-1 or PD-L1) that is expressed at a lower level on a Treg than CTLA4.

35. The therapeutic combination of claim 25, wherein the anti-ICM antigen binding molecule antagonizes an ICM (e.g., PD-1 or PD-L1) that is expressed at a higher level on an immune effector cell (e.g., an effector T cell, macrophage, dendritic cell, B cell, etc.) than on a Treg.

36. The therapeutic combination of claim 27, wherein the anti-PD-1 antigen binding molecule is a MAb, non-limiting examples of which include nivolumab, pembrolizumab, pidilizumab, and MEDI-0680 (AMP-514), AMP-224, JSOOl-PD-1, SHR- 1210, Gendor PD-1, PDR001, CT-011, REGN2810, BGB-317 or an antigen-binding fragment thereof, or one that competes with nivolumab, pembrolizumab, pidilizumab, or MEDI-0680 for binding to PD-1.

37. The therapeutic combination of claim 27 or claim 36, wherein the anti-PD-1 antigen-binding molecule binds specifically to one or more amino acids of the amino acid sequence set forth in SEQ ID NO:9 (/'.e., residues 62 to 86 of the native PD-1 sequence set forth in SEQ ID NO: 10), SEQ ID NO: 11 (/.e., residues 118 to 136 of the native PD-1 sequence set forth in SEQ ID NO: 10) and SEQ ID NO: 12 (/'.e.,

corresponding to residue 66 to 97 of the native PD-1 sequence set forth in SEQ ID NO: 10).

38. The therapeutic combination of claim 28, wherein the anti-PD-Ll antigen-binding molecule is a MAb, non-limiting examples of which include durvalumab (MEDI4736), atezolizumab (Tecentriq), avelumab, BMS-936559/MDX-1105,

MSB0010718C, LY3300054, CA-170, GNS-1480 and MPDL3280A, or an antigen-binding fragment thereof.

39. The therapeutic combination of claim 28 or claim 38, wherein the anti-PD-Ll antigen-binding molecule binds specifically to one or more amino acids in the amino acid sequence set forth in SEQ ID NO: 13 (/'.e., residues 279 to 290 of the full length native PD-L1 amino acid sequence set forth in SEQ ID NO: 14), is one that competes with any one of durvalumab (MEDI4736), atezolizumab (Tecentriq), avelumab, BMS-936559/MDX-1105, MSB0010718C, LY3300054, CA-170, GNS-1480 and MPDL3280A for binding to PD-L1.

40. The therapeutic combination of claim 29, wherein the anti-CTLA4 antigen-binding molecule is a MAb, representative examples of which include ipilimumab and tremelimumab, or an antigen-binding fragment thereof, or is one that competes with ipilimumab or tremelimumab for binding to CTLA4.

41. The therapeutic combination of claim 29 or claim 40, wherein the anti-CTLA4 antigen-binding molecule binds specifically to one or more amino acids in an amino acid sequence selected from the sequences set forth in any one of SEQ ID NO: 15 (/'.e., residues 25 to 42 of the full-length native CTLA4 amino acid sequence set forth in

SEQ ID NO: 16), SEQ ID NO: 17 (/'.e., residues 43 to 65 of the native CTLA4 sequence set forth in SEQ ID NO: 16), and SEQ ID NO: 18 (/'.e., residues 96 to 109 of the native CTLA4 sequence set forth in SEQ ID NO: 16).

42. The therapeutic combination of any one of claim 22 to 41, wherein the RANK or ICM antigen-binding molecule is linked to an immunoglobulin constant chain

(e.g., an IgGl, IgG2a, IgG2b, IgG3, or IgG4 constant chain).

43. The therapeutic combination of claim 42, wherein the immunoglobulin constant chain may comprise a light chain selected from a k light chain or l light chain; and a heavy chain selected from a yl heavy chain, y2 heavy chain, y3 heavy chain, and y4 heavy chain.

44. The therapeutic combination of any one of claim 22 to 43, wherein the therapeutic combination comprises, consists or consists essentially of the RANK antagonist antigen-binding molecule of any one of claims 1 to 4 and two or more different anti-ICM antigen-binding molecules.

45. The therapeutic combination of claim 44, wherein the therapeutic combination comprises, consists or consists essentially of a RANK antagonist antigen binding molecule described herein and at least two of an anti-CTLA4 antigen-binding molecule, an anti-PD-1 antigen-binding molecule and an anti-PD-Ll antigen-binding molecule.

46. The therapeutic combination of any one of claim 22 to 45, wherein individual antigen-binding molecules of the therapeutic combination are in the form of discrete components.

47. The therapeutic combination of any one of claim 22 to 45, wherein individual antigen-binding molecules of the therapeutic combination are fused or otherwise conjugated (either directly or indirectly) to one another.

48. The therapeutic combination of claim 47, wherein the therapeutic combination is in the form of a multispecific antagonist agent, comprising the RANK antagonist antigen-binding molecule of any one of claims 1 to 4 and the at least one anti-ICM antigen-binding molecule.

49. The therapeutic combination of claim 48, wherein the multispecific agent is a complex of two or more polypeptides.

50. The therapeutic combination of claim 48, wherein the multispecific agent is a single chain polypeptide.

51. The therapeutic combination of claim 50, wherein the RANK antagonist antigen-binding molecule is conjugated to the N-terminus or to the C-terminus of an individual anti-ICM antigen-binding molecule.

52. The therapeutic combination of claim 50 or claim 51, wherein RANK antagonist antigen-binding molecule and anti-ICM antigen-binding molecule are connected directly or by an intervening linker (e.g., a polypeptide linker).

53. The therapeutic combination of any one of claims 22 to 52, wherein the therapeutic combination is contained in a delivery vehicle (e.g., a liposome, a nanoparticle, a microparticle, a dendrimer or a cyclodextrin).

54. A multispecific antigen-binding molecule for co-antagonizing RANK and at least one ICM, the multispecific antigen-binding molecules comprising, consisting or consisting essentially of the RANK antagonist antigen-binding molecule of any one of claims 1 to 4 and at least one anti-ICM antigen-binding molecule.

55. The multispecific antigen-binding molecule of claim 54, wherein the RANK antagonist antigen-binding molecule is an antibody or antigen-binding fragment thereof that binds specifically to and antagonizes RANK.

56. The multispecific antigen-binding molecule of claim 54 or claim 55, wherein individual anti-ICM antigen-binding molecules are selected from antibodies or antigen-binding fragments that binds specifically to and antagonize a corresponding ICM.

57. The multispecific antigen-binding molecule of claim 55 or claim 56, wherein the antibody and/or antigen-binding fragments are connected directly or by an intervening linker (e.g., a chemical linker or a polypeptide linker).

58. The multispecific antigen-binding molecule of any one of claims 54 to 57, which is in the form of a single chain polypeptide in which the RANK antagonist antigen binding molecule and at least one anti-ICM antigen-binding molecule are operably connected.

59. The multispecific antigen-binding molecule of any one of claims 54 to 57, which is in the form of discrete polypeptide chains in which the RANK antagonist antigen-binding molecule and at least one anti-ICM antigen-binding molecule are linked to or otherwise associated with one another to form a complex.

60. The multispecific antigen-binding molecule of any one of claims 54 to 59, which is bivalent, trivalent, or tetravalent.

61. The multispecific antigen-binding molecule of any one of claims 54 to 60, wherein the RANK antagonist antigen-binding molecule and the at least one anti-ICM antigen-binding molecule are selected from Fab, Fab', F(ab')2, and Fv molecules and complementarity determining regions (CDRs).

62. The multispecific antigen-binding molecule of any one of claims 54 to 61, wherein individual antigen-binding molecules comprise a constant domain that is independently selected from the group consisting of IgG, IgM, IgD, IgA, and IgE.

63. The multispecific antigen-binding molecule of any one of claims 54 to 62, which comprises a tandem scFv (taFv or scFv2), diabody, dAb2/VHH2, knobs-in-holes derivative, Seedcod-IgG, heteroFc-scFv, Fab-scFv, scFv-Jun/Fos, Fab'-Jun/Fos, tribody,

DNL-F(ab)3, SCFV -CH1/CL, Fab-scFv2, IgG-scFab, IgG-scFv, scFv-IgG, scFv2-Fc, F(ab')2-SCFV2, SCDB-FC, scDb-CH3, Db-Fc, scFv2-H/L, DVD-Ig, tandAb, scFv-dhlx-scFv, dAb2-IgG, dAb-IgG, dAb-Fc-dAb, tandab, DART, BiKE, TriKE, ITIFC-VH, crosslinked MAbs, Cross MAbs, MAb2, FIT-Ig, electrostatically matched antibodies, symmetric IgG-like antibodies, LUZ-Y, Fab-exchanged antibodies, or a combination thereof.

64. The multispecific antigen-binding molecule of any one of claims 54 to 63, wherein individual antigen-binding molecules are linked to an immunoglobulin constant chain (e.g., IgGl, IgG2a, IgG2b, IgG3, and IgG4).

65. The multispecific antigen-binding molecule of claim 64, wherein the immunoglobulin constant chain comprises a light chain selected from a k light chain and l light chain, and/or a heavy chain selected from a yl heavy chain, y2 heavy chain, y3 heavy chain, and y4 heavy chain.

66. The multispecific antigen-binding molecule of any one of claims 54 to 65, wherein the multispecific antigen-binding molecule antagonizes PD-1, and the anti-PD-1 antigen-binding molecule (e.g., an antibody or antigen-binding fragment thereof) binds specifically to one or more amino acids of an amino acid sequence selected from SEQ ID NO:9 (/.e., residues 62 to 86 of the native human PD-1 sequence set forth in SEQ ID NO: 10), SEQ ID NO: 11 (/.e., residues 118 to 136 of the native human PD-1 sequence set forth in SEQ ID NO: 10) and SEQ ID NO: 12 (/'.e., corresponding to residue 66 to 97 of the native human PD-1 sequence set forth in SEQ ID NO: 10).

67. The multispecific antigen-binding molecule of claim 66, wherein the anti-PD-1 antigen-binding molecule (e.g., an antibody or antigen-binding fragment thereof) comprises a heavy chain and a light chain of a MAb selected from nivolumab, pembrolizumab, pidilizumab, and MEDI-0680 (AMP-514), AMP-224, JSOOl-PD-1, SHR-1210, Gendor PD-1, PDR001, CT-011, REGN2810, BGB-317 or antigen-binding fragments thereof.

68. The multispecific antigen-binding molecule of any one of claims 54 to 67, wherein the multispecific antigen-binding molecule antagonizes PD-L1, and the anti-PD- L1 antigen-binding molecule (e.g., an antibody or antigen-binding fragment thereof) binds specifically to one or more amino acids of the amino acid sequence set forth in SEQ ID NO: 13 (/.e., residues 279 to 290 of the native human PD-L1 amino acid sequence as set forth in SEQ ID NO: 14).

69. The multispecific antigen-binding molecule of claim 68, wherein the anti- PD-L1 antigen-binding molecule (e.g., an antibody or antigen-binding fragment thereof) comprise a heavy chain and a light chain of a MAb selected from durvalumab

(MEDI4736), atezolizumab (Tecentriq), avelumab, BMS-936559/MDX-1105,

MSB0010718C, LY3300054, CA-170, GNS-1480 and MPDL3280A, or antigen-binding fragments thereof.

70. The multispecific antigen-binding molecule of any one of claims 54 to 69, wherein the multispecific antigen-binding molecule antagonizes CTLA4, and the anti-CTLA4 antigen-binding molecule (e.g., an antibody or antigen-binding fragment thereof) binds specifically to one or more amino acids of an amino acid sequence selected from SEQ ID NO: 15 (/'.e., residues 25 to 42 of the full-length native PD-CTLA4 amino acid sequence set forth in SEQ ID NO: 16), SEQ ID NO: 17 (/'.e., residues 43 to 65 of the native CTLA4 sequence set forth in SEQ ID NO: 16), and SEQ ID NO: 18 (/'.e., residues 96 to 109 of the native CTLA4 sequence set forth in SEQ ID NO: 16).

71. The multispecific antigen-binding molecule of claim 70, wherein the anti-CTLA4 antigen-binding molecule (e.g., an antibody or antigen-binding fragment thereof) comprises a heavy chain and a light chain of a MAb selected from ipilimumab and tremelimumab, or antigen-binding fragments thereof.

72. The multispecific antigen-binding molecule of any one of claims 54 to 71, wherein the multispecific antigen-binding molecule comprises, consists or consists essentially of the RANK antagonist antigen-binding molecule of any one of claims 1 to 4 and an anti-PD-1 antigen-binding molecule.

73. The multispecific antigen-binding molecule of any one of claims 54 to 71, wherein the multispecific antigen-binding molecule comprises, consists or consists essentially of the RANK antagonist antigen-binding molecule of any one of claims 1 to 4 and an anti-PD-Ll antigen-binding molecule.

74. The multispecific antigen-binding molecule of any one of claims 54 to 71, wherein the multispecific antigen-binding molecule comprises, consists or consists

essentially of the RANK antagonist antigen-binding molecule of any one of claims 1 to 4, an anti-PD-1 antigen-binding molecule and an anti-PD-Ll antigen-binding molecule.

75. The multispecific antigen-binding molecule of any one of claims 54 to 71, wherein the multispecific antigen-binding molecule comprises, consists or consists essentially of the RANK antagonist antigen-binding molecule of any one of claims 1 to 4, an anti-PD-1 antigen-binding molecule and an anti-CTLA4 antigen-binding molecule.

76. The multispecific antigen-binding molecule of any one of claims 54 to 71, wherein the multispecific antigen-binding molecule comprises, consists or consists essentially of the RANK antagonist antigen-binding molecule of any one of claims 1 to 4, and an anti-PD-Ll antigen-binding molecule.

77. A method of producing the therapeutic combination of any one of claims 54 to 76, the method comprising combining the RANK antagonist antigen-binding molecule of any one of claims 1 to 4, and at least one anti-ICM antigen-binding molecule to thereby produce the therapeutic combination.

78. The method of claim 77, further comprising generating an antigen binding molecule that binds specifically to and antagonizes a target polypeptide (e.g., RANK or an ICM) of the therapeutic combination (e.g., by immunizing an animal with an immunizing polypeptide comprising an amino acid sequence corresponding to an the target polypeptide; and identifying and/or isolating a B cell from the animal, which binds specifically to the target polypeptide or at least one region thereof; and producing the antigen-binding molecule expressed by that B cell).

79. The method of claim 77 or claim 78, further comprising derivatizing the antigen-binding molecule so generated to produce a derivative antigen-binding molecule with the same epitope-binding specificity as the antigen-binding molecule.

80. The method of claim 79, wherein the derivative antigen-binding molecule is selected from antibody fragments, illustrative examples of which include Fab, Fab', F(ab' , Fv, single chain (scFv), one-arm and domain antibodies (including, for example, shark and camelid antibodies), and fusion proteins comprising an antibody, and any other modified configuration of an immunoglobulin molecule that comprises an antigen binding/ recognition site.

81. A construct that comprise a nucleic acid sequence encoding the multispecific antigen-binding molecule of any one of claims 54 to 76, in operable connection with one or more control sequences.

82. The construct of claim 81, which in the form of an expression construct, representative examples of which include plasmids, cosmids, phages, and viruses.

83. A host cell that contain the construct of claim 81 or claim 82.

84. A pharmaceutical composition comprising the therapeutic combination of any one of claims 22 to 53, or the multispecific antigen-binding molecule of any one of claims 54 to 76, and a pharmaceutically acceptable carrier.

85. The composition of claim 84, further comprising at least one ancillary agent selected from a chemotherapeutic agent (e.g., selected from

antiproliferative/antineoplastic drugs, cytostatic agents, agents that inhibit cancer cell invasion, inhibitors of growth factor function, anti-angiogenic agents, vascular damaging

- Ill -

agents, etc.), or an immunotherapeutic agent {e.g., cytokines, cytokine-expressing cells, antibodies, etc.).

86. A method for stimulating or augmenting immunity in a subject, the method comprising, consisting or consisting essentially of administering to the subject an effective amount of the therapeutic combination of any one of claims 22 to 53, or the multispecific antigen-binding molecule of any one of claims 54 to 76, to thereby stimulate or augment immunity in the subject.

87. The method of claim 86, wherein the RANK antagonist antigen-binding molecule and the at least one anti-ICM antigen-binding molecule of the therapeutic combination are provided as discrete components, and the components are administered concurrently to the subject.

88. The method of claim 87, wherein the RANK antagonist antigen-binding molecule is administered simultaneously with the at least one anti-ICM antigen-binding molecule.

89. The method of claim 87, wherein the RANK antagonist antigen-binding molecule and the at least one anti-ICM antigen-binding molecule are administered sequentially.

90. The method of claim 89, wherein the RANK antagonist antigen-binding molecule is administered prior to administration of the at least one anti-ICM antigen binding molecule.

91. The method of claim 89, wherein the RANK antagonist antigen-binding molecule is administered after administration of the at least one anti-ICM antigen binding molecule.

92. The method of any one of claims 86 to 91, wherein the stimulated or augmented immunity comprises a beneficial host immune response, illustrative examples of which include any one or more of the following : reduction in tumor size; reduction in tumor burden; stabilization of disease; production of antibodies against an endogenous or exogenous antigen; induction of the immune system; induction of one or more components of the immune system; cell-mediated immunity and the molecules involved in its production; humoral immunity and the molecules involved in its production; antibody-dependent cellular cytotoxicity (ADCC) immunity and the molecules involved in its production; complement-mediated cytotoxicity (CDC) immunity and the molecules involved in its production; natural killer cells; cytokines and chemokines and the molecules and cells involved in their production; antibody-dependent cytotoxicity; complement-dependent cytotoxicity; natural killer cell activity; and antigen-enhanced cytotoxicity.

93. The method of claim 92, wherein the stimulated or augmented immunity includes a pro-inflammatory immune response.

94. A method for inhibiting the development or progression of

immunosuppression or tolerance to a tumor in a subject, the method comprising, consisting or consisting essentially of contacting the tumor with the therapeutic combination of any one of claims 22 to 53, or the multispecific antigen-binding molecule of any one of claims 54 to 76, to thereby inhibit the development or progression of immunosuppression or tolerance to the tumor in the subject.

95. The method of claim 94, wherein the therapeutic combination or multispecific antigen-binding molecule also contacts an antigen-presenting cell (e.g., a dendritic cell) that presents a tumor antigen to the immune system.

96. A method for inhibiting the development, progression or recurrence of a cancer in a subject, the method comprising, consisting or consisting essentially of administering to the subject an effective amount of the therapeutic combination of any one of claims 22 to 53, or the multispecific antigen-binding molecule of any one of claims 54 to 76, to thereby inhibit the development, progression or recurrence the cancer in the subject.

97. A method for treating a cancer in a subject, the method comprising, consisting or consisting essentially of administering to the subject an effective amount of the therapeutic combination of any one of claims 22 to 53, or the multispecific antigen binding molecule of any one of claims 54 to 76, to thereby treat the cancer.

98. The method of claim 97, wherein the cancer is selected from melanoma, breast cancer, colon cancer, ovarian cancer, endometrial and uterine carcinoma, gastric or stomach cancer, pancreatic cancer, prostate cancer, salivary gland cancer, lung cancer, hepatocellular cancer, glioblastoma, cervical cancer, liver cancer, bladder cancer, hepatoma, rectal cancer, colorectal cancer, kidney cancer, vulval cancer, thyroid cancer, hepatic carcinoma, anal carcinoma, penile carcinoma, testicular cancer, esophageal cancer, tumors of the biliary tract, head and neck cancer, and squamous cell carcinoma. In some particular embodiments, the cancer is a metastatic cancer.

99. The method of any one of claims 86 to 98, wherein the subject has reduced or impaired responsiveness to immunomodulatory agents, for example a subject that has reduced or impaired responsiveness to ICM molecule antagonists (e.g. , an anti-PD-1 or anti-PD-Ll immunotherapy).

100. The method of any one of claims 86 to 99, further comprising concurrently administering to the subject an effective amount of an ancillary anti-cancer agent.

101. The method of claim 100, wherein the ancillary anti-cancer agent includes a chemotherapeutic agent, external beam radiation, a targeted radioisotope, and a signal transduction inhibitor.

102. A kit for stimulating or augmenting immunity, for inhibiting the development or progression of immunosuppression or tolerance to a tumor, or for treating a cancer in a subject, the kit comprising any one or more of the therapeutic combination of any one of claims 22 to 53, or the multispecific antigen-binding molecule of any one of claims 54 to 76.